The rockefeller university

Recent work reveals that the innate immune system is able to recognize self-targets and initiate an inflammatory response similar to that of pathogens. One novel example of this innate autoimmunity is ischemia/reperfusion (I/R) injury. in which reperfusion of the ischemic tissues elicits an acute inflammatory response activated by natural IgM (nIgM) binding to ischemia-specific self-antigens, which are non-muscle myosin heavy chains type II (NMHC-II) subtype A and C Subsequently, the complement lectin pathway is activated and eventually tissue injury occurs. Although earlier studies in the intestinal model showed that the classical complement pathway did not initiate I/R injury, C1q deposition was still observed in the local injured tissues by imaging analysis Moreover, the involvement of the alternative complement pathway became unclear due to conflicting reports using different knockout mice To explore the immediate downstream pathway following nIgM-ischemic antigen interaction, we isolated the nIgM-ischemic antigen immunocomplexes from the local tissue of animals treated in the intestinal I/R injury model, and examined the presence of initial molecules of three complement pathways. Our results showed that mannan-binding lectin (MBL), the early molecule of the lectin pathway, was present in the nIgM-ischemic Ag immunocomplex In addition, C1q, the initial molecule of the classical pathway was also detected on the immunocomplex However, Factor B, the early molecule in the alternative pathway, was not detected in the immunocomplex. To further examine the role of the alternative pathway in I/R injury, we utilized Factor B knockout mice in the intestinal model Our results showed that Factor B knockout mice were not protected from local tissue injury, and their complement system was activated in the local tissues by nIgM during I/R These results indicated that the lectin complement pathway operates immediately downstream of the nIgM-ischemic antigen interaction during intestinal I/R. Furthermore. the classical complement pathway also appears to interact with the of nIgM-ischemic antigen immunocomplex Finally, the alternative complement pathway is not involved in I/R injury induction in the current intestinal model (C) 2009 Elsevier Ltd All rights reserved

The bacterial RNA polymeras holoenzyme consists of a catalytic core enzyme in complex with a sigma factor that is required for promoter-specific transcription initiation. During initiation, members of the sigma(70) family of sigma factors contact two conserved promoter elements, the -10 and -35 elements, which are separated by similar to 17 base pairs (bp). sigma(70) family members contain four flexibly linked domains. Two of these domains, sigma(2) and sigma(4), contain determinants for interactions with the promoter -10 and -35 elements respectively. sigma(2) and sigma(4) also contain core-binding determinants. When bound to core the inter-domain distance between sigma(2) and sigma(4) matches the distance between promoter elements separated by similar to 17 bp. Prior work indicates that during early elongation the nascent RNA-assisted displacement of sigma(4) from core can enable the holoenzyme to adopt a configuration in which sigma(2) and sigma(4) are bound to 'promoter-like' DNA elements separated by a single base pair. Here we demonstrate that holoenzyme can also adopt configurations in which sigma(2) and sigma(4) are bound to 'promoter-like' DNA elements separated by 0, 2 or 3 bp. Thus, our findings suggest that displacement of sigma(4) from core enables the RNA polymerase holoenzyme to adopt a broad range of 'elongation-specific' configurations.

Most patients with autoimmune polyendocrine syndrome type I (APS-I) display chronic mucocutaneous candidiasis (CMC). We hypothesized that this CMC might result from autoimmunity to interleukin (IL)-17 cytokines. We found high titers of autoantibodies (auto-Abs) against IL-17A, IL-17F, and/or IL-22 in the sera of all 33 patients tested, as detected by multiplex particle-based flow cytometry. The auto-Abs against IL-17A, IL-17F, and IL-22 were specific in the five patients tested, as shown by Western blotting. The auto-Abs against IL-17A were neutralizing in the only patient tested, as shown by bioassays of IL-17A activity. None of the 37 healthy controls and none of the 103 patients with other autoimmune disorders tested had such auto-Abs. None of the patients with APS-I had auto-Abs against cytokines previously shown to cause other well-defined clinical syndromes in other patients (IL-6, interferon [IFN]-gamma, or granulocyte/macrophage colony-stimulating factor) or against other cytokines (IL-1 beta, IL-10, IL-12, IL-18, IL-21, IL-23, IL-26, IFN-beta, tumor necrosis factor [alpha], or transforming growth factor beta). These findings suggest that auto-Abs against IL-17A, IL-17F, and IL-22 may cause CMC in patients with APS-I.

We present three measurements of the top-quark mass in the lepton plus jets channel with approximately 1.9 fb(-1) of integrated luminosity collected with the CDF II detector using quantities with minimal dependence on the jet energy scale. One measurement exploits the transverse decay length of b-tagged jets to determine a top-quark mass of 166.9(-8.5)(+9.5)(stat) +/- 2.9(syst) GeV/c(2), and another the transverse momentum of electrons and muons from W-boson decays to determine a top-quark mass of 173.5(-8.9)(+8.8)(stat) +/- 3.8(syst) GeV/c(2). These quantities are combined in a third, simultaneous mass measurement to determine a top-quark mass of 170.7 +/- 6.3(stat) +/- 2.6(syst) GeV/c(2) .

Lens epithelium-derived growth factor (LEDGF) fusion proteins can direct HIV-1 DNA integration to novel sites in the host genome. The C terminus of LEDGF contains an integrase binding domain (IBD), and the N terminus binds chromatin. LEDGF normally directs integrations to the bodies of expressed genes. Replacing the N terminus of LEDGF with chromatin binding domains (CBDs) from other proteins changes the specificity of HIV-1 DNA integration. We chose two well-characterized CBDs: the plant homeodomain (PHD) finger from ING2 and the chromodomain from heterochromatin binding protein 1 alpha (HP1 alpha). The ING2 PHD finger binds H3K4me3, a histone mark that is associated with the transcriptional start sites of expressed genes. The HP1 alpha chromodomain binds H3K9me2,3, histone marks that are widely distributed throughout the genome. A fusion protein in which the ING2 PHD finger was linked to the LEDGF IBD directed integrations near the start sites of expressed genes. A similar fusion protein in which the HP1 alpha chromodomain was linked to the LEDGF IBD directed integrations to sites that differed from both the PHD finger fusion-directed and LEDGF-directed integration sites. The ability to redirect HIV-1 DNA integration may help solve the problems associated with the activation of oncogenes when retroviruses are used in gene therapy.

To combat variola virus in bioterrorist attacks, it is desirable to develop a noninvasive vaccine. Based on the vaccinia Tiantan (VTT) strain, which was historically used to eradicate the smallpox in China, we generated a modified VTT (MVITZCI) by removing the hemagglutinin gene and an 11,944 bp genomic region from HindIII fragment C2L to F3L. MVTTZCI was characterized for its host cell range in vitro and preclinical safety and efficacy profiles in mice. Despite replication-competency in some cell lines, unlike VTT, MVTTZCI did not cause death after intracranial injection or body weight loss after intranasal inoculation. MVTTZCI did not replicate in mouse brain and was safe in immunodeficient mice. MVTTZCI induced neutralizing antibodies via the intranasal route of immunization. One time intranasal immunization protected animals from the challenge of the pathogenic vaccinia WR strain. This study established proof-of-concept that the attenuated replicating MVTTZCI may serve as a safe noninvasive smallpox vaccine candidate. (C) 2009 Elsevier Ltd. All rights reserved.

From the earliest work in our laboratory, we hypothesized, and with studies conducted in both clinical research and animal models, we have shown that drugs of abuse, administered or self-administered, on a chronic basis, profoundly alter stress-responsive systems. Alterations of expression of specific genes involved in stress responsivity, with increases or decreases in mRNA levels, receptor, and neuropeptide levels, and resultant changes in hormone levels, have been documented to occur after chronic intermittent exposure to heroin, morphine, other opiates, cocaine, other stimulants, and alcohol in animal models and in human molecular genetics. The best studied of the stress-responsive systems in humans and mammalian species in general is undoubtedly the HPA axis. In addition, there are stress-responsive systems in other parts in the brain itself, and some of these include components of the HPA axis, such as CRF and CRF receptors, along with POMC gene and gene products. Several other stress-responsive systems are known to influence the HPA axis, such as the vasopressin-vasopressin receptor system. Orexin-hypocretin, acting at its receptors, may effect changes which suggest that it should be properly categorized as a stress-responsive system. However, less is known about the interactions and connectivity of some of these different neuropeptide and receptor systems, and in particular, about the possible connectivity of fast-acting (e.g., glutamate and GABA) and slow-acting (including dopamine, serotonin, and norepinephrine) neurotransmitters with each of these stress-responsive components and the resultant impact, especially in the setting of chronic exposure to drugs of abuse. Several of these stress-responsive systems and components, primarily based on our laboratory-based and human molecular genetics research of addictive diseases, will be briefly discussed in this review. (C) 2009 Elsevier B.V. All rights reserved.

Calcium signals are involved in a large variety of physiological processes. Their versatility relies on the diversity of spatio-temporal behaviors that the calcium concentration can display. Calcium entry through inositol 1,4,5-trisphosphate (IP(3)) receptors (IP(3)R's) is a key component that participates in both local signals such as "puffs'' and in global waves. IP(3)R's are usually organized in clusters on the membrane of the endoplasmic reticulum and their spatial distribution has important effects on the resulting signal. Recent high resolution observations [1] of Ca(2+) puffs offer a window to study intra-cluster organization. The experiments give the distribution of the number of IP(3)R's that open during each puff without much processing. Here we present a simple model with which we interpret the experimental distribution in terms of two stochastic processes: IP(3) binding and unbinding and Ca(2+)-mediated inter-channel coupling. Depending on the parameters of the system, the distribution may be dominated by one or the other process. The transition between both extreme cases is similar to a percolation process. We show how, from an analysis of the experimental distribution, information can be obtained on the relative weight of the two processes. The largest distance over which Ca(2+)-mediated coupling acts and the density of IP(3)-bound IP(3)R's of the cluster can also be estimated. The approach allows us to infer properties of the interactions among the channels of the cluster from statistical information on their emergent collective behavior.

Papers in this special issue of Physiology and Behavior illustrate some of the roles of steroid and thyroid hormones in achieving a modern version of what physiologists have described as 'the unity of the body'. Included are papers on stress physiology and on endocrine disruptors, studied at several levels. (C) 2009 Published by Elsevier Inc.

A total of 91 university community participants were evaluated on the five personality factors (NEO-Five Factor Inventory; NEO-FFI) and with the Conners Continuous Performance Test (CPT). Gender differences for the Conners CPT variables were found such that the men were faster and less variable in their reaction times than the women, and the women were more sensitive in terms of signal detection and conservative in their responses than the men, consistent with the data reported by Conners, Epstein, Angold, and Klaric (2003) for a normative sample of 9-18 year olds. Interesting relationships between personality factors and the CPT variables were seen for the male and female groups, especially in terms of response style and vigilance. For the combined genders, higher vigilance was associated with higher neuroticism, lower extraversion, and lower agreeableness. Lower conscientiousness was associated with a more risk-taking/ less conservative response bias. The men were more vigilant than the women, and for the men, this higher vigilance was related to lower conscientiousness, with trends for relationships with greater neuroticism and lower agreeableness. In the women, higher vigilance was related to lower extraversion, consistent with other studies. Men who were more extraverted also tended to be less conservative/more risk taking in their response style. A more risk-taking response style in women was associated with higher neuroticism, higher openness, and lower conscientiousness. The findings are interpreted in terms of underlying biological bases of behavior.